Flow cytometric detection of proteolysis in peptide libraries synthesised on optically encoded supports

The concept of optically encoding particles for solid phase organic synthesis has existed in the literature for several years. However, there remains a significant challenge to producing particles that are capable of withstanding harsh solvents and reagents whilst maintaining the integrity and range...

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Bibliographic Details
Published in:Molecular bioSystems 2008-01, Vol.4 (7), p.774-778
Main Authors: Surawski, Peter P T, Battersby, Bronwyn J, Lawrie, Gwendolyn A, Ford, Kym, Rühmann, Andreas, Marcon, Lionel, Kozak, Darby, Trau, Matt
Format: Article
Language:English
Subjects:
Flow Cytometry - methods
Fluorescent Dyes - chemistry
Hydrolysis
Microspheres
Organosilicon Compounds - chemistry
Peptide Hydrolases - metabolism
Peptide Library
Peptides - chemical synthesis
Peptides - chemistry
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Summary:The concept of optically encoding particles for solid phase organic synthesis has existed in the literature for several years. However, there remains a significant challenge to producing particles that are capable of withstanding harsh solvents and reagents whilst maintaining the integrity and range of the optical encoding. In this study, a new generation of fluorescently encoded support particles was used for both solid phase peptide synthesis and on-particle analysis of proteolysis in a multiplexed, flow cytometric assay. The success of the assay was demonstrated through the use of a model protease, trypsin. Our results show that the use of solid supports with high peptide yield, high swellability in water and high penetration of the enzyme into the interior of the particle is not absolutely necessary for proteolysis assays.
ISSN:1742-206X
1742-2051
DOI:10.1039/b718513h